The long-term product goal of this project is a small molecule therapeutic to prevent or treat Type 1 diabetes (T1D). Our objective is to reduce the inflammatory response triggered by the pro-inflammatory cytokine macrophage migration inhibitory factor (MIF). Under physiological conditions MIF is secreted by ? cells and regulates their function, but during immunoinflammatory events the pancreatic islets secrete high amounts of MIF that mediate islet apoptosis. Immunoneutralization or genetic deletion of MIF in mouse models of T1D has correlated with halting the progression of T1D or completely preventing the disease. Since MIF is an upstream regulator of the inflammatory cascade, small molecule therapeutics targeting MIF activity are expected to provide effective treatment for T1D, which currently afflicts 3 million people in the US alone and for which there is no curative therapy. To this end, we have identified novel small molecule MIF inhibitors. Our lead proprietary small molecules, 2F3 and 1A5, block MIF-driven cellular activation pathways that are associated with the immunopathology of T1D, including apoptosis and production of proinflammatory mediators. 2F3 and 1A5 are minimally cytotoxic and are structurally unique, possessing functional groups that have not been previously associated with MIF inhibitory activity. The immediate objectives of the Phase I is to demonstrate the therapeutic efficacy of L2's proprietary 2F3 and 1A5 small molecules to reverse or prevent T1D in the well established T1D non-obese diabetic (NOD) mouse model. NOD mice, which are the mainstay of preclinical diabetes research as they share similarities with human T1D, will be treated with our lead compounds and controls in a time and dose dependent manner. Therapeutic efficacy will be monitored by measuring blood glucose levels and ? cell death by our recently developed novel droplet digital PCR. In addition we will perform cellular assays measuring Th1, Th2 and T-regulatory cytokine levels, as well as apoptosis, to determine the effects of the 2F3 and 1A5 on MIF-driven inflammatory process. All of these efforts are expected to yield a lead compound suitable for further development towards a small molecule therapeutic for T1D.